Influence of Liver Fibrosis on Pien Tze Huang by RNA-Seq Analysis of Circular RNAs

Purpose: We investigate the regulation circRNAs in the damaged fibrotic liver treated with Pien Tze Huang (PZH) by RNA-seq. Methods: Liver tissue circRNAs expression profiles from 8-week carbon tetrachloride (CCl4)-induced, and PZH-treated and CCl4-induced liver fibrosis control mice were investigated by RNA-seq and dysregulated circRNAs in PZH-treated were identified. Various bioinformatics analyses established competing endogenous RNA (ceNRA) in circRNAs-miRNAs-mRNAs axis. Moreover, GO annotation and Kegg pathway were analyzed to identify the relational biological processes and pathway. qRT–PCR validation was performed using LX-2 cell sample pairs and SYBR Green assays. Results: We identified 91 differentially expressed circRNAs in the PZH-treated compared with control mice, of which 58 up-regulated circRNAs and 43 down-regulated circRNAs. The ceRNA network were constructed suggesting the potential role of circRNAs is the regulation of target gene expression as a miRNAs sponge. Moreover, GO and Kegg pathway analysis showed that these predicted mRNAs were related to liver fibrosis-related pathways such as TNF, PI3K-Akt, and MAPK signaling pathway. Meanwhile, the results of validation qRT-PCR of target genes in LX-2 cells were in accordance with RNA-seq with high confidence. Conclusions: We identified the dysregulated circRNAs and established functional network of circRNA-miRNA-mRNA to further reveal the potential interactions and biological processes in PZH-treated liver fibrosis mice. These findings indicate that mmu_circ_Slc25a25 may modulate the transcription of certain genes, such as Slc7a11 and Slc39a14, which may provide a more precise and comprehensive perspective for clarifying the role of PZH therapy as a potential agent for treatment of liver fibrosis. Investigation of dysregulated circRNA in liver tissue of 8-week PZH-treated or without mice by RNA-seq, using Illumina Hiseq 4000.